US11035779B2 - Particle container and particle filling apparatus - Google Patents
Particle container and particle filling apparatus Download PDFInfo
- Publication number
- US11035779B2 US11035779B2 US16/712,281 US201916712281A US11035779B2 US 11035779 B2 US11035779 B2 US 11035779B2 US 201916712281 A US201916712281 A US 201916712281A US 11035779 B2 US11035779 B2 US 11035779B2
- Authority
- US
- United States
- Prior art keywords
- particle
- region
- particles
- measurement
- width
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active, expires
Links
- 239000002245 particle Substances 0.000 title claims abstract description 219
- 238000005259 measurement Methods 0.000 claims abstract description 147
- 230000008859 change Effects 0.000 claims abstract description 14
- 230000004308 accommodation Effects 0.000 claims description 66
- 239000007788 liquid Substances 0.000 claims description 47
- 125000006850 spacer group Chemical group 0.000 claims description 16
- 239000000463 material Substances 0.000 claims description 14
- 230000002093 peripheral effect Effects 0.000 claims description 7
- 230000032258 transport Effects 0.000 description 16
- 239000012528 membrane Substances 0.000 description 8
- 230000005484 gravity Effects 0.000 description 7
- 238000003752 polymerase chain reaction Methods 0.000 description 6
- 238000005558 fluorometry Methods 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 4
- 239000011347 resin Substances 0.000 description 4
- 229920005989 resin Polymers 0.000 description 4
- 238000012360 testing method Methods 0.000 description 3
- 101100010147 Oryza sativa subsp. japonica DOF1 gene Proteins 0.000 description 2
- 238000001746 injection moulding Methods 0.000 description 2
- 238000009434 installation Methods 0.000 description 2
- 230000005389 magnetism Effects 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 229920000515 polycarbonate Polymers 0.000 description 2
- 239000004417 polycarbonate Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 239000004925 Acrylic resin Substances 0.000 description 1
- 229920000178 Acrylic resin Polymers 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- NBVXSUQYWXRMNV-UHFFFAOYSA-N fluoromethane Chemical compound FC NBVXSUQYWXRMNV-UHFFFAOYSA-N 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 235000015250 liver sausages Nutrition 0.000 description 1
- 239000006249 magnetic particle Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000005373 porous glass Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000010349 pulsation Effects 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000012508 resin bead Substances 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1484—Optical investigation techniques, e.g. flow cytometry microstructural devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
- B01L3/502784—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1404—Handling flow, e.g. hydrodynamic focusing
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1456—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/0627—Sensor or part of a sensor is integrated
- B01L2300/0654—Lenses; Optical fibres
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0848—Specific forms of parts of containers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0877—Flow chambers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/12—Specific details about materials
- B01L2300/123—Flexible; Elastomeric
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1404—Handling flow, e.g. hydrodynamic focusing
- G01N2015/1418—Eliminating clogging of debris
Definitions
- the present disclosure generally relates to a particle container and a particle filling apparatus.
- a testing (measuring) apparatus is available for performing fluorescence observation (fluorometry) or the like to evaluate a sample (particles) containing DNA or protein to be observed (measured).
- the sample is heated and reacted in a cartridge (particle container) of the testing apparatus.
- the width of the gap may be reduced to a size approximately equal to the diameter of the particles, which thereby prevents the particles from being piled up in the particle-measuring direction.
- a smaller width of the gap prolongs the time required for filling the particles, which poses another problem.
- the present disclosure provides a particle container that includes a measurement section in which a measurement region is formed so as to measurably accommodate multiple particles, in which the time required for filling particles into the measurement region can be reduced.
- the present disclosure provides a particle container including a measurement section in which a measurement region is formed and the measurement section measurably accommodates multiple particles in the measurement region.
- the measurement section is configured to change a width of the measurement region in the measuring direction of measuring the particles.
- FIG. 1A is a perspective view illustrating an external appearance of a cartridge (particle container) according to an embodiment
- FIG. 1B is a cross-sectional view of the cartridge.
- FIGS. 2A to 2F are cross-sectional views illustrating states of in which the gap width of a measurement region changes.
- FIGS. 3A to 3D are cross-sectional views illustrating states in which the width of measurement region changes.
- FIG. 4 is a general view illustrating a measurement system according to the embodiment, which, for example, fills spherical samples (minute particles).
- FIGS. 5A to 5D are cross-sectional views illustrating types of transport units and their operations according to the embodiment.
- FIGS. 6A to 6D are views for explaining problems to be solved with the embodiment, in which FIGS. 6A and 6B are cross-sectional views and FIGS. 6C and 6D each include an enlarged cross-sectional view and an enlarged top view.
- FIGS. 7A to 7D are views illustrating types of width changing units and their operations according to the embodiment, in which FIGS. 7A and 7B are cross-sectional views, FIG. 7C is a perspective view, and FIG. 7D is a cross-sectional view.
- FIGS. 8A to 8C are cross-sectional views illustrating an operation example of the transport unit and the width changing unit according to the embodiment.
- FIGS. 9A to 9D are cross-sectional views illustrating operation examples of the width changing unit according to the embodiment and also illustrating examples of width changing units that enable the operation.
- FIGS. 10A to 10C are cross-sectional views illustrating operation examples of the width changing unit according to the embodiment and also illustrating an example of a particle container that enables the operation.
- FIGS. 11A to 11C are views for explaining about the diameter of a spherical sample and the change of the width, in which FIGS. 11A and 11B each include an enlarged cross-sectional view and an enlarged top view and FIG. 11C is an enlarged cross-sectional view.
- FIG. 12 is a cross-sectional view illustrating a particle container according to a second embodiment.
- the present disclosure relates to a particle container (a cartridge) that accommodates spherical samples (particles) in a preliminary region (accommodation region) formed therein and that fills the spherical samples into an observation region (measurement region) of which the gap width in a measuring direction (particle observation direction) is set at a value related to the diameter of a particle.
- the present disclosure relates to a particle filling apparatus for filling particles into the measurement region of the particle container for a short period of time and also relates to a measurement system that includes the particle filling apparatus.
- FIGS. 1A, 1B and 4 An example of a particle container and an example of a particle filling apparatus (and a measurement system including the particle filling apparatus) according to the present disclosure will be described with reference to FIGS. 1A, 1B and 4 .
- Reference 001 denotes a particle to be observed.
- Reference 002 denotes a measurement region in which particles 001 are placed during observation, in other words, the particles 001 are measurably accommodated.
- the measurement region 002 is formed between an upper plate 003 and a lower plate 004 , which serve as plates that oppose each other, and the measurement region 002 is configured such that the gap of the measurement region 002 is changeable by using an external force.
- Reference 008 denotes a side portion disposed between the upper plate 003 and the lower plate 004 .
- a measurement section in which the measurement region 002 is formed is constituted at least by the upper plate 003 , the lower plate 004 , and the side portion 008 .
- Reference 005 denotes an accommodation region that accommodates particles 001 and is disposed next to the measurement region 002 so as to be in communication with the measurement region 002 .
- the accommodation region 005 is configured to have a maximum height (a maximum width in the measuring direction) being at least greater than a maximum height of the measurement region 002 .
- An accommodation section in which the accommodation region 005 is formed is constituted at least by the lower plate 004 , a junction portion 009 (described later), and an opening-formed plate 030 (described later).
- the width of the accommodation region 005 in the measuring direction of measuring the particles 001 may be twice or more as large as the diameter of the particle 001 .
- the accommodation section includes a liquid-droplet generation portion 007 , which will be described later.
- Reference 009 denotes a junction portion that is disposed between the accommodation region 005 and the upper plate 003 or the lower plate 004 .
- Reference 006 denotes a sample inlet that allows a liquid sample to enter the accommodation region 005 .
- Reference 007 denotes a liquid-droplet generation portion by which a liquid sample is transformed into particles 001 .
- Reference 030 denotes an opening-formed plate in which the sample inlet 006 and an opening 012 (described later) are formed.
- the accommodation region 005 accommodates multiple particles 001 .
- a liquid sample to be transformed into particles for observation is poured into the accommodation region 005 through the sample inlet 006 .
- the sample inlet 006 can have a cover to prevent sample contamination.
- the liquid sample poured is subsequently transformed into minute liquid droplets, which are particles 001 , by the liquid-droplet generation portion 007 .
- the liquid-droplet generation portion 007 may be formed of a porous glass membrane, an emulsification membrane having micro holes, or microchannels for generating liquid droplets.
- the wall of the accommodation region 005 to be in contact with a heat source can have a structure having a high thermal conductivity.
- the floor plate in the accommodation region 005 may be made of a metal plate or a thin resin plate.
- the continuous phase may generate gas bubbles.
- the gas bubbles which disturb observation of the particles 001 , are desirably prevented from entering the measurement region 002 . Accordingly, the height of the accommodation region 005 is made greater than a maximum height of the measurement region 002 , and gas bubbles generated in the accommodation region 005 are thereby retained in the accommodation region 005 .
- Providing a space in the accommodation region 005 for storing entrapped gas bubbles is also effective in preventing gas bubbles from entering the measurement region 002 .
- the measurement region 002 is formed between the upper plate 003 and the lower plate 004 .
- the side portion 008 is provided between the upper plate 003 and the lower plate 004 , thereby forming a container (measurement section) having the measurement region formed therein.
- the side portion 008 defines the width of the gap (i.e., the width between the upper plate 003 and the lower plate 004 in the measuring direction).
- the measurement region 002 is positioned adjacent to the accommodation region 005 and configured to take the particles 001 stored in the accommodation region 005 into the measurement region 002 .
- the width of the gap between the upper plate 003 and the lower plate 004 can be changed by an external force. A mechanism for this will be described below with reference to FIGS. 2A to 2F .
- FIG. 2A is a cross section of the measurement section that forms the measurement region 002 to be filled with particles 001 .
- FIGS. 2B and 2C are cross-sectional views illustrating states in which an external force is applied to the measurement section of FIG. 2A .
- the upper plate 003 has flexibility, and deformation of the upper plate 003 narrows the gap of the measurement region 002 .
- the side portion 008 has flexibility, and deformation of the side portion 008 narrows the gap of the measurement region 002 .
- the deformation is elastic.
- the structures and materials of the upper plate 003 , the lower plate 004 , and the side portion 008 are determined so as to deform in an expected manner. Note that when no external force is applied, the gap of the measurement region 002 is stably maintained as illustrated in FIG. 2A . It is also necessary to determine the structures and materials of the upper plate 003 and the side portion 008 accordingly.
- the side portion 008 and the junction portion 009 that connect the upper plate 003 to other structures are made of a flexible material, and a first spacer 010 is disposed in the measurement region 002 that is the gap between the upper plate 003 and the lower plate 004 .
- the first spacer 010 is made of a cushion material and defines the height of the upper pate 003 .
- the first spacer 010 can be made of a rubber material, but compatibility with the continuous phase needs to be considered. Especially fluorocarbon rubber is compatible with various types of continuous phases.
- the second spacer 011 is disposed inside the measurement region 002 that is the gap between the first plate 003 and the second plate 004 . Accordingly, the material of the second spacer 011 is such that the second spacer 011 does not deform by the external force and is not degraded by the continuous phase. Note that in FIG. 2F , the second spacer 011 is disposed on the lower plate 004 , but the second spacer may be disposed on the upper plate 003 .
- the second spacer 011 is desirably shaped as a structure having a sharp portion as illustrated in FIG. 2F .
- the sharp portion can move particles 001 easily away from the second spacer 011 when the gap is narrowed, which prevents the particles 001 from being crushed.
- Particles 001 are a target object of optical measurement, such as fluorescence observation (fluorometry), in the measurement region 002 .
- at least one of the upper plate 003 and the lower plate 004 is made of a light-transmissive material.
- the light-transmissive material to be used may be glass, quartz, or a resin material, such as acrylic resin or polycarbonate.
- a resin can be used in the case of the material being subjected to deformation due to an external force.
- the area of the upper plate 003 where the width of the gap is constant can be large after the deformation of the measurement region 002 .
- Devising the shape of the upper plate 003 leads to expansion of area of the upper plate 003 where the width of the gap is constant after the measurement region 002 deforms.
- the thickness of the upper plate 003 in the measuring direction may be changed depending on positions on a surface of the upper plate 003 (on a surface orthogonally intersecting the measuring direction). More specifically, a peripheral portion of the upper plate 003 is made thinner than a central portion of the upper plate 003 .
- FIG. 3A the thickness of the upper plate 003 in the measuring direction may be changed depending on positions on a surface of the upper plate 003 (on a surface orthogonally intersecting the measuring direction). More specifically, a peripheral portion of the upper plate 003 is made thinner than a central portion of the upper plate 003 .
- a groove that deforms preferentially is formed in a circumferential portion of the upper plate 003 .
- Different thickness portions may be provided only in the lower plate 004 , instead of the upper plate 003 , or, as illustrated in FIGS. 3C and 3D , may be provided both in the upper plate 003 and in the lower plate 004 .
- width changing units (which will be described in detail later) may be provided to apply loads to the upper plate 003 and the lower plate 004 , respectively.
- the width changing unit may be used to displace one of the upper plate 003 and the lower plate 004 , and a different device other than the width changing unit may be used to displace the other one of the plates.
- the different device may be a member that has a certain thickness and is in contact with the other one of the plates.
- the other one of the plates can also receive a load as a reaction force from the member. This can eliminate the necessity of providing a plurality of the width changing units and can simplify the device for applying load, which leads to a reduction in the manufacturing cost of the particle filling apparatus and also in the maintenance and operation cost.
- the thicknesses of the upper plate 003 and the lower plate 004 may be appropriately determined so as to bear loads to be received. The thicknesses may be the same or may be different from each other.
- the gap width of the measurement region 002 can become substantially constant at the central portion thereof.
- a particle filling apparatus and a measurement system 200 that includes the particle filling apparatus will be described with reference to FIG. 4 .
- Reference 013 is a holder base onto which a particle container is set and fixed.
- Reference 014 is a thermoregulator that changes the temperature of spherical samples in the particle container.
- Reference 015 is a pump to be used for transforming the liquid sample contained in the sample inlet portion into liquid droplets at the liquid-droplet generation portion 007 .
- Reference 016 is a transport unit for transporting particles 001 placed in the accommodation region 005 toward the measurement region 002 .
- Reference 017 is a width changing unit that changes the width of the gap of the measurement region 002 .
- Reference 018 is a camera for observation of particles 001 disposed in the measurement region 002 .
- Reference 019 is a light source to be used for emitting light to the particles 001 when the particles 001 are observed (measured) by using the camera 018 . These devices are actuated by a control unit 020 .
- the particle container is fixed onto the holder base 013 .
- the particle container is subsequently filled with a liquid that serves as a continuous phase. Note that the particle container may be filled with the continuous phase in advance.
- a liquid sample to be measured is poured into the sample inlet 006 .
- the pump 015 is connected to the opening 012 and actuated.
- particles 001 are generated in the form of minute liquid droplets at the liquid-droplet generation portion 007 and stored in the accommodation region 005 .
- a syringe pump may be used as the pump 015 by attaching a syringe to the opening 012 .
- a syringe pump may be used as the pump 015 by attaching a syringe to the opening 012 .
- a non-pulsation pump can generate liquid droplets of less variation in size.
- the particles 001 can be heated by the thermoregulator 014 .
- the polymerase chain reaction (PCR) can be carried out by subjecting the particles 001 to a temperature cycle between 60° C. and 90° C.
- a device that can regulate temperature such as a Peltier element, can be used for the thermoregulator 014 .
- thermoregulator 014 is disposed on the floor plate in the accommodation region 005 in FIG. 4 . However, the thermoregulator 014 may be disposed in the measurement region 002 . This configuration is suitable for observation of the particles 001 in the condition of temperature change.
- thermoregulator 014 may be disposed both in the accommodation region 005 and in the measurement region 002 and may be selected suitably according to an application objective.
- the thermoregulator 014 in the measurement region may be capable of controlling temperature more precisely than the thermoregulator 014 in the accommodation region 005 .
- the particle filling apparatus at least includes the width changing unit 017 .
- the particle filling apparatus may include the transport unit 016 .
- the transport unit 016 transports the particles 001 toward the measurement region 002 , and the width changing unit 017 changes the gap of the measurement region 002 . This process will be described later.
- the light source 019 emits light to the particles 001 placed in the measurement region 002 , and the camera 018 takes an image of the particles 001 .
- light emitted by the light source 019 may be, for example, white light for taking an image of particles' external appearances or ultraviolet light for fluorescence observation (fluorometry), to be selected depending on an application objective.
- the camera 018 and the light source 019 are disposed above the upper plate 003 .
- the position is not limited to this.
- a plurality of the light sources 019 and the cameras 018 may be provided.
- control unit 020 These devices are controlled by the control unit 020 so as to automatically carry out a series of processing from pouring of the liquid sample.
- FIGS. 5A, 5B, and 5C illustrate an example in which a device that can incline the particle container is employed for the transport unit 016 .
- FIG. 5B illustrates an example in which the specific gravity of the particles 001 is greater than that of the continuous phase
- FIG. 5C illustrates an example in which the specific gravity of the particles 001 is smaller than that of the continuous phase.
- the particles 001 can move toward the measurement region 002 by gravity. Heating of the thermoregulator 014 may generate gas bubbles in the particle container.
- the gas bubbles which disturb observation of the particles 001 using the camera 018 , may be prevented from remaining in the measurement region 002 .
- the gas bubbles can be treated when the transport unit 016 is actuated.
- the accommodation region 005 is raised to a higher position as illustrated in FIG. 5B , thereby moving the gas bubbles toward the accommodation region 005 .
- the accommodation region 005 can be raised temporarily as illustrated in FIG. 5B , which causes the gas bubbles to move into the accommodation region 005 and causes the gas bubbles to be trapped in the opening 012 or a hollow such as the sample inlet 006 . Consequently, the accommodation region 005 is lowered as illustrated in FIG. 5C , which causes the particles 001 to move to the measurement region 002 .
- it is also effective to subject the particle container to an impact or vibrations so as to facilitate movement of the gas bubbles from the measurement region 002 .
- FIG. 5D illustrate an example in which a device that can move a magnet 021 between the accommodation region 005 and the measurement region 002 is employed for the transport unit 016 .
- the particles 001 are moved using magnetism. Accordingly, the particles 001 need to be transportable by magnetism. For example, in the case of the liquid droplet, it is effective to mix minute magnetic particles in the liquid sample in advance.
- the rotation device for the transport unit 016 , and the rotation device rotates the holder base 013 , which produces centrifugal forces and thereby moves the particles 001 toward the measurement region 002 .
- the specific gravity of the particles 001 is larger, the gas bubbles and the particles 001 move in directions opposite to each other, which facilitates movement of the gas bubbles from the measurement region 002 toward the accommodation region 005 .
- the particles 001 can move in the measurement region 002 easily, which is advantageous because the particles 001 can be filled into the measurement region 002 quickly ( FIG. 6A ).
- the particles 001 may pile up in the measurement region 002 , causing some particles 001 not to be observed from the camera 018 ( FIG. 6C ).
- This problem can be solved by limiting the width of the gap to a level corresponding to the diameter of each particle 001 and by arranging the particles 001 in one layer ( FIG. 6D ).
- the width changing unit 017 operates to avoid such a problem.
- the transport unit 016 is actuated when the gap is wide as illustrated in FIG. 6A and thereby causes the particles 001 to move quickly into the measurement region 002 .
- FIG. 7A illustrates a state of the measurement region 002 after the particles 001 are transported.
- a single axis actuator is used to press the width changing unit 017 against the upper plate 003 and thereby narrow the gap. This causes the particles 001 to be arranged in one layer.
- a linear stage may be used as the single axis actuator.
- the width changing unit 017 is not limited to the single axis actuator insofar as the width of the gap can be changed.
- the measurement region 002 of which the gap is narrowed in advance is provided.
- the particles 001 is moved into the measurement region 002 by actuating the transport unit 016 .
- the gap is widened by actuating a gap control unit.
- the particles 001 collected at the entrance of the measurement region 002 can be thereby taken into the measurement region 002 .
- the measurement region 002 can be filled with the particles 001 quickly, and gas bubbles generated in the accommodation region 005 during heating can be prevented from entering the measurement region 002 .
- the width changing unit 017 it is also effective to devise a method of controlling the width changing unit 017 so as to arrange liquid droplets in one layer smoothly.
- the gap width is changed after the measurement region 002 is filled with the particles 001 as is the example illustrated in FIGS. 7A to 7D
- the gap is narrowed progressively from the lowest portion of the measurement region 002 in FIG. 9A .
- the particles 001 can be thereby arranged in one layer in the measurement region 002 without the particles 001 being clogged.
- the gap is widened to take in the particles 001
- the gap is widened progressively from a portion of the measurement region 002 near the accommodation region 005 as illustrated in FIG. 9B .
- the particles 001 can be thereby taken smoothly into the measurement region 002 .
- multiple actuators may be provided for the width changing unit 017 as illustrated in FIG. 9C .
- the multiple actuators apply pressure on a portion of the measurement region 002 near the accommodation region 005 and on a deeper portion of the measurement region 002 at different timings.
- the contact portion of the width changing unit 017 has a shape projecting toward a portion of the upper plate 003 located deeper in the measurement region 002 .
- the upper plate 003 is pressed progressively from the deeper portion toward the accommodation region 005 .
- the width changing unit 017 directly presses and deforms the upper plate 003 (or the peripheral portion of the upper plate 003 ) that defines the measurement region 002 .
- the width changing unit 017 is not limited to this configuration.
- a negative pressure or a positive pressure is applied in the particle container in advance.
- the width changing unit 017 releases the pressure inside the particle container, which can change the gap of the measurement region 002 .
- particles 001 are stored in the accommodation region 005 .
- the upper plate 003 is deformed due to the pump 015 or the like applying a positive pressure or a negative pressure.
- the airtight particle container is maintained with the gap being changed.
- the particle container is shaped as illustrated in FIG. 10A when a positive pressure is applied inside the particle container, and the particle container is shaped as illustrated in FIG. 10B when a negative pressure is applied.
- the particle container includes a headspace portion 024 communicating with the accommodation region 005 and a release plug 023 that plugs the headspace portion 024 and the accommodation region 005 .
- the width changing unit 017 actuates the release plug 023 , the pressure in the particle container is released, thereby causing the upper plate 003 to return to its original shape and change the gap.
- FIG. 10C illustrates an opening state of the release plug 023 .
- the headspace portion 024 has the opening 012 that is formed at the particle container and has an edge.
- the headspace portion 024 also has a plug that is movable upward and downward and is disposed so as to plug the opening 012 .
- the plug includes a projection 025 for opening the release plug 023 and a stopper 026 that stops movement of the projection 025 so as to prevent the projection 025 from opening the release plug 023 during normal operation.
- the plug has a sealing member that prevents the continuous phase from releasing out of the particle container when the release plug 023 is open.
- any type of device can be employed in the gap control unit insofar as the device can release the stopper 026 and opens the release plug 023 .
- a linear type motor can be used.
- the width of the gap after the gap is changed is desirably set at a value approximately equal to or more than the diameter of each particle 001 .
- the width of the gap being 1.66 times more than the diameter, as illustrated in FIG. 11A , at least a one half of the diameter portion or more of a particle 001 can be observed from the camera 018 even if the particles 001 are stacked. Fluorescence observation (fluorometry) can be carried out if such an amount of portion of each particle 001 is observable (measurable).
- the fluorescence observation can be still carried out even if a less portion of each particle 001 than the above is observable in a case in which the diameter of each particle 001 is large and the camera 018 is capable of large magnification.
- the width of the gap exceeds 1.83 times of the diameter, a central 10% portion of the diameter of a particle 001 can be still imaged. If the width of the gap is twice the diameter or more, the central portion of a particle 001 may not be imaged when the particles 001 are stacked. Accordingly, the width of the gap can be equal to or less than twice the diameter.
- each particle 001 In the case of observing the external appearance of each particle 001 , it is necessary to observe outlines of upper particles 001 and lower particles 001 . As illustrated in FIG. 11C , when the depth of field of the camera 018 is DOF1, the width of the gap need to be reduced to the diameter+DOF1 or less.
- All the particles 001 in the particle container may not necessarily be in the same size.
- the diameter of the particles 001 can be defined as a maximum diameter of particles 001 of a central 95% portion in the size distribution of the particles 001 contained in the particle container.
- the width of the gap can be prescribed with respect to the particle diameter in accordance with an application objective, which enables the camera 018 to obtain information of the particles 001 in line with the objective.
- FIG. 1 schematically illustrates a configuration of a particle container according to the present embodiment. All the components of the particle container are made of polycarbonate except for the liquid-droplet generation portion 007 . Sheet members and members manufactured using injection molding are assembled using an adhesive or a joining technology. In the present embodiment, the floor plate of the accommodation region 005 and the lower plate 004 are formed of one sheet member, and other portions of the housing are manufactured using injection molding. Note that a 0.2 mm thick sheet is adopted to facilitate heat conduction.
- a porous membrane made of silicon by using photo-processing is used as the liquid-droplet generation portion 007 .
- the porous membrane has a large number of equally shaped through-holes, and the surface of the porous membrane is subjected to hydrophobic treatment.
- the porous membrane is disposed so as to cover the sample inlet 006 and adhered thereto.
- the sheet member and the member to which the porous membrane is adhered are joined to each other so as to form a container.
- Threads for connection with various connectors are formed at the sample inlet 006 and the opening 012 , which enables connection with the pump 015 and sealing by using a lid.
- each hole of the porous membrane is adjusted so as to be able to produce liquid droplets of approximately 100 ⁇ m in diameter when the sample passes through the holes.
- the height of the accommodation region 005 is designed to be 500 ⁇ m and the height (i.e., width) of the measurement region 002 is designed to be 400 ⁇ m.
- FIG. 4 is a diagram schematically illustrating a configuration of a filling system of particles 001 according to the present embodiment.
- the holder base 013 is formed of a metal plate, and the particle container can be fixed thereto.
- the holder base 013 is equipped with a rotation shaft 022 that enables the holder base 013 to incline.
- the particle container fixed to the holder base 013 is filled with an oil that serves as the continuous phase.
- a syringe is installed in the opening 012 , and the syringe is connected to a syringe pump, which serves as the pump 015 .
- An aqueous liquid sample is poured into the sample inlet 006 , and then the syringe pump is actuated. This causes the liquid sample to pass through the liquid-droplet generation portion 007 . As a result, a large number of liquid droplets are generated in the accommodation region 005 .
- the syringe is removed after the generation of the liquid droplets, and the particle container is sealed with lids covering the opening 012 and the sample inlet 006 .
- the syringe may be utilized in place of a lid. It is effective to automate installation of the syringe, connection or disconnection of the pump 015 , pouring of the liquid sample, and installation of a lid. After the particle container is sealed, the liquid droplets are heated by actuating the Peltier element that serves as the thermoregulator 014 . Note that in the case of performing PCR, the liquid droplets are subjected several times to a temperature cycle between 60° C. and 90° C.
- the holder base 013 is inclined by actuating a linear type motor that serves as the transport unit 016 .
- the specific gravity of the liquid sample is greater than that of the oil in this case, the liquid droplets move toward the measurement region 002 by inclining the holder base 013 .
- the diameter of each liquid droplet is 100 ⁇ m, whereas the width of the gap of the measurement region 002 is 400 ⁇ m, which is large enough to take in the liquid droplets quickly.
- the gap of the measurement region 002 is narrowed by actuating the linear type motor that serves as the width changing unit 017 and by deforming the upper plate 003 .
- the contact portion of the width changing unit 017 to be in contact with the upper plate 003 is shaped so as to have a hollow at the center as illustrated in FIGS. 7C and 7D .
- the hollow region becomes an observation region.
- the gap actuator unit is controlled to cause the width of the gap in the hollow region to be 266 mm or less, which is 1.66 times greater than the diameter of each liquid droplet. As a result, the liquid droplets are arranged in one layer in the observation region.
- the light source 019 emits ultraviolet light, and the fluorescence observation is performed using the camera 018 . Thus, all the liquid droplets present within the observation region can be observed.
- multiple particles 001 to be observed are put in the particle container that can change the width of gap of the measurement region 002 .
- the width of gap of the measurement region 002 is changed approximately to the diameter of each particle 001 . This enables the particles 001 to be arranged in one layer in a short period of time.
- FIG. 12 schematically illustrates a configuration of a particle container according to the present embodiment.
- a large number of dents each having a size similar to the diameter of each liquid droplet are formed in a central portion of the upper plate 003 .
- the dents open toward the inside of the particle container.
- the width changing unit 017 deforms the upper plate 003 and thereby causes liquid droplets to enter the dents and to be settled therein.
- the width of gap of the measurement region 002 after the gap is changed can be set such that the distance between the protrusions of the upper plate 003 and the lower plate 004 is smaller than the diameter of each liquid droplet. Movement of each liquid droplet is thereby prevented by the dent, which facilitates identification of each liquid droplet during observation using the camera 018 .
- using the above-described cartridge and system enables the particles 001 to be filled quickly into the measurement region 002 in one layer and also enables the particles 001 to be prevented from moving.
- a particle container that includes a measurement section in which a measurement region is formed so as to measurably accommodate multiple particles, the time required for filling multiple particles into the measurement region can be reduced.
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Dispersion Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Hematology (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Fluid Mechanics (AREA)
- Sampling And Sample Adjustment (AREA)
- Optical Measuring Cells (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Description
Claims (18)
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JPJP2018-243764 | 2018-12-26 | ||
JP2018-243764 | 2018-12-26 | ||
JP2018243764 | 2018-12-26 |
Publications (2)
Publication Number | Publication Date |
---|---|
US20200209142A1 US20200209142A1 (en) | 2020-07-02 |
US11035779B2 true US11035779B2 (en) | 2021-06-15 |
Family
ID=71124097
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/712,281 Active 2039-12-23 US11035779B2 (en) | 2018-12-26 | 2019-12-12 | Particle container and particle filling apparatus |
Country Status (2)
Country | Link |
---|---|
US (1) | US11035779B2 (en) |
JP (1) | JP2020103283A (en) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4290997A (en) * | 1978-02-28 | 1981-09-22 | Kommandiittiyhtio Finnpipette Osmo A. Suovaniemi | Apparatus for automatic measurement of the results of agglutination tests |
WO2008146754A1 (en) | 2007-05-23 | 2008-12-04 | Trust Co., Ltd. | Container for liquid reaction mixture, reaction-promoting device using the same and method therefor |
US20190054466A1 (en) * | 2017-08-17 | 2019-02-21 | Abbott Point Of Care Inc. | Single-use test device for imaging blood cells |
US20200102587A1 (en) * | 2017-04-05 | 2020-04-02 | Hitachi High-Technologies Corporation | Nucleic acid amplification method and nucleic acid analyzer |
-
2019
- 2019-12-12 US US16/712,281 patent/US11035779B2/en active Active
- 2019-12-20 JP JP2019230524A patent/JP2020103283A/en active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4290997A (en) * | 1978-02-28 | 1981-09-22 | Kommandiittiyhtio Finnpipette Osmo A. Suovaniemi | Apparatus for automatic measurement of the results of agglutination tests |
WO2008146754A1 (en) | 2007-05-23 | 2008-12-04 | Trust Co., Ltd. | Container for liquid reaction mixture, reaction-promoting device using the same and method therefor |
US20200102587A1 (en) * | 2017-04-05 | 2020-04-02 | Hitachi High-Technologies Corporation | Nucleic acid amplification method and nucleic acid analyzer |
US20190054466A1 (en) * | 2017-08-17 | 2019-02-21 | Abbott Point Of Care Inc. | Single-use test device for imaging blood cells |
Also Published As
Publication number | Publication date |
---|---|
JP2020103283A (en) | 2020-07-09 |
US20200209142A1 (en) | 2020-07-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN111587149B (en) | Digital microfluidic device and method of use thereof | |
US9671365B2 (en) | Electrowetting dispensing devices and related methods | |
US6989234B2 (en) | Method and apparatus for non-contact electrostatic actuation of droplets | |
KR100865105B1 (en) | Microfabricated elastomeric valve and pump systems | |
US8790595B2 (en) | Apparatus and methods for microfluidic applications | |
Teste et al. | Selective handling of droplets in a microfluidic device using magnetic rails | |
CN112469504A (en) | Control of evaporation in digital microfluidics | |
WO2011002957A2 (en) | Droplet actuator devices and methods | |
CN114904594A (en) | Liquid bead separation in microfluidics | |
EP2944940B1 (en) | Aid for filling liquid, and method for filling liquid | |
CN106104271A (en) | There is micro-fluid chip and the manufacture thereof in conical bead trapping chamber | |
Lagoy et al. | Microfluidic devices for behavioral analysis, microscopy, and neuronal imaging in Caenorhabditis elegans | |
US20100086444A1 (en) | Biochip manufacturing method and biochip manufacturing device | |
JP2007232719A (en) | Ceramic microarray spotting device for biological assay printing (ceramic plate and dispensing method for dispensing liquid) | |
US11035779B2 (en) | Particle container and particle filling apparatus | |
Compera et al. | Upscaling of pneumatic membrane valves for the integration of 3D cell cultures on chip | |
KR20060080585A (en) | Microfluidic packaging | |
US20050069949A1 (en) | Microfabricated Fluidic Structures | |
JP2009115732A (en) | Micro-inspection chip, method for micro-inspection chip to determine quantity of a liquid, and inspection method | |
KR101718491B1 (en) | Liquid patterning apparatus | |
JP2020051574A (en) | Sheet-like gasket | |
CN114669336B (en) | Micro-droplet generation method | |
de Gruiter et al. | Department of Precision and Microsystems Engineering | |
US20200156077A1 (en) | A sample cartridge for incubating and/or analyzing a dispersion of particles, cells or droplets | |
Xu | Droplet-based microfluidic system for multiple-droplet trapping, storing, and clustering |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
FEPP | Fee payment procedure |
Free format text: ENTITY STATUS SET TO UNDISCOUNTED (ORIGINAL EVENT CODE: BIG.); ENTITY STATUS OF PATENT OWNER: LARGE ENTITY |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: SENT TO CLASSIFICATION CONTRACTOR |
|
AS | Assignment |
Owner name: CANON KABUSHIKI KAISHA, JAPAN Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:EBISAWA, HISAFUMI;REEL/FRAME:052093/0442 Effective date: 20191203 |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NOTICE OF ALLOWANCE MAILED -- APPLICATION RECEIVED IN OFFICE OF PUBLICATIONS |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: PUBLICATIONS -- ISSUE FEE PAYMENT RECEIVED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: PUBLICATIONS -- ISSUE FEE PAYMENT VERIFIED |
|
STCF | Information on status: patent grant |
Free format text: PATENTED CASE |